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用于在转染的成纤维细胞和淋巴细胞中稳定和瞬时表达HLA - DR的高效cDNA表达载体。

Efficient cDNA expression vectors for stable and transient expression of HLA-DR in transfected fibroblast and lymphoid cells.

作者信息

Long E O, Rosen-Bronson S, Karp D R, Malnati M, Sekaly R P, Jaraquemada D

机构信息

Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.

出版信息

Hum Immunol. 1991 Aug;31(4):229-35. doi: 10.1016/0198-8859(91)90092-n.

Abstract

cDNA expression vectors with several useful features were constructed. First, the long terminal repeat of Rous sarcoma virus was used as a promoter to obtain high levels of expression in various cells of human and mouse origin. Second, cis-linked expression units that confer resistance either to mycophenolic acid or the neomycin analog G418 were inserted to facitate the isolation of transfected cells expressing the cDNA of interest. Third, by replicating in simian COS cells, these vectors can be used for efficient transient expression. cDNA fragments encoding the DR alpha or DR beta chains of human class II major histocompatibility complex antigens were inserted into these vectors and high levels of cell surface HLA-DR antigen were obtained after cotransfection into mouse and human fibroblasts. These vectors were also successfully used to correct the inability of a class II-negative B cell line, derived from a patient with a congenital immunodeficiency, to present peptide antigen to DR-restricted T cells.

摘要

构建了具有多种有用特性的cDNA表达载体。首先,劳斯肉瘤病毒的长末端重复序列被用作启动子,以在人和小鼠来源的各种细胞中获得高水平的表达。其次,插入赋予对霉酚酸或新霉素类似物G418抗性的顺式连接表达单元,以促进表达感兴趣的cDNA的转染细胞的分离。第三,通过在猴COS细胞中复制,这些载体可用于高效瞬时表达。将编码人II类主要组织相容性复合体抗原的DRα或DRβ链的cDNA片段插入这些载体中,并在共转染到小鼠和人成纤维细胞后获得高水平的细胞表面HLA-DR抗原。这些载体还成功地用于纠正源自先天性免疫缺陷患者的II类阴性B细胞系向DR限制性T细胞呈递肽抗原的无能。

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